New hard tick (Acari: Ixodidae) reports and detection of Rickettsia in ticks from Sierra Nevada de Santa Marta, Colombia.

The Sierra Nevada de Santa Marta (SNSM), located in northern Colombia, is considered a geographical island with high levels of biodiversity and endemism. However, little is known about tick species and their associated microorganisms at the SNSM. In this study we sampled host-seeking ticks in areas of the town of Minca within the SNSM. We collected 47 ticks identified as Amblyomma pacae, Amblyomma longirostre, Amblyomma ovale, Amblyomma mixtum, Haemaphysalis juxtakochi, Ixodes sp. cf. Ixodes affinis and Ixodes sp. Of these ticks, we tested for Rickettsia spp. by amplifying the gltA, SCA1, and 16S rRNA genes via PCR. Rickettsia amblyommatis was detected in one pool of 3 larvae and in a female of A. pacae. Additonally, we isolated Rickettsia sp. belonging to the group of spotted fevers in larvae of A. longirostre. This study reports new findings of six species of ticks and two species of Rickettsia within the SNSM.

New insights into the molecular phylogeny, biogeographical history, and diversification of Amblyomma ticks (Acari: Ixodidae) based on mitogenomes and nuclear sequences.

BACKGROUND: Amblyomma is the third most diversified genus of Ixodidae that is distributed across the Indomalayan, Afrotropical, Australasian (IAA), Nearctic and Neotropical biogeographic ecoregions, reaching in the Neotropic its highest diversity. There have been hints in previously published phylogenetic trees from mitochondrial genome, nuclear rRNA, from combinations of both and morphology that the Australasian Amblyomma or the Australasian Amblyomma plus the Amblyomma species from the southern cone of South America, might be sister-group to the Amblyomma of the rest of the world. However, a stable phylogenetic framework of Amblyomma for a better understanding of the biogeographic patterns underpinning its diversification is lacking. METHODS: We used genomic techniques to sequence complete and nearly complete mitochondrial genomes -ca. 15 kbp- as well as the nuclear ribosomal cluster -ca. 8 kbp- for 17 Amblyomma ticks in order to study the phylogeny and biogeographic pattern of the genus Amblyomma, with particular emphasis on the Neotropical region. The new genomic information generated here together with genomic information available on 43 ticks (22 other Amblyomma species and 21 other hard ticks-as outgroup-) were used to perform probabilistic methods of phylogenetic and biogeographic inferences and time-tree estimation using biogeographic dates. RESULTS: In the present paper, we present the strongest evidence yet that Australasian Amblyomma may indeed be the sister-group to the Amblyomma of the rest of the world (species that occur mainly in the Neotropical and Afrotropical zoogeographic regions). Our results showed that all Amblyomma subgenera (Cernyomma, Anastosiella, Xiphiastor, Adenopleura, Aponomma and Dermiomma) are not monophyletic, except for Walkeriana and Amblyomma. Likewise, our best biogeographic scenario supports the origin of Amblyomma and its posterior diversification in the southern hemisphere at 47.8 and 36.8 Mya, respectively. This diversification could be associated with the end of the connection of Australasia and Neotropical ecoregions by the Antarctic land bridge. Also, the biogeographic analyses let us see the colonization patterns of some neotropical Amblyomma species to the Nearctic. CONCLUSIONS: We found strong evidence that the main theater of diversification of Amblyomma was the southern hemisphere, potentially driven by the Antarctic Bridge's intermittent connection in the late Eocene. In addition, the subgeneric classification of Amblyomma lacks evolutionary support. Future studies using denser taxonomic sampling may lead to new findings on the phylogenetic relationships and biogeographic history of Amblyomma genus.

Case report: Diagnosis and autogenous vaccine treatment of herpesvirus in a green turtle (Chelonia mydas) in Santa Marta, Colombia.

This study reports the first case of fibropapillomatosis (FP) in the green turtle Chelonia mydas that has been successfully diagnosed and treated in Colombia. Worldwide, FP has reached epizootic proportions as it has been reported in marine turtles of tropical and subtropical waters, and in severe cases, it reduces the probability of survival. Treatment has been elusive as multiple surgical excisions are needed due to tumor recurrence. In this case, one green turtle with multiple tumors was diagnosed by histopathology and molecular detection of the chelonid herpesvirus 5 (ChHV5) by means of amplification and sequencing of the DNA polymerase (DNApol) gene. Two separate treatments that consisted of autogenous vaccines and surgical excisions were applied; the first one had a partial success as one out of the tumors treated reappeared after 3 months post-treatment. Treatment 2 consisted of an autogenous vaccine enriched with adjuvants and applied at increasing doses, after which, the tumor significatively decreased in size and was surgically removed. At the end of the 6 months follow-up period, no tumor recurrence was observed, and the turtle was in apparent optimal health conditions. These findings, although limited, suggest a possible treatment that might help to contain this epizootic problem.

A Comparative Phylogeography of Three Marine Species with Different PLD Modes Reveals Two Genetic Breaks across the Southern Caribbean Sea.

The comparative phylogeography of marine species with contrasting dispersal potential across the southern Caribbean Sea was evaluated by the presence of two putative barriers: the Magdalena River plume (MRP) and the combination of the absence of a rocky bottom and the almost permanent upwelling in the La Guajira Peninsula (ARB + PUG). Three species with varying biological and ecological characteristics (i.e., dispersal potentials) that inhabit shallow rocky bottoms were selected: Cittarium pica (PLD < 6 days), Acanthemblemaria rivasi (PLD < 22 days), and Nerita tessellata (PLD > 60 days). We generated a set of SNPs for the three species using the ddRad-seq technique. Samples of each species were collected in five locations from Capurganá to La Guajira. For the first time, evidence of a phylogeographic break caused by the MRP is provided, mainly for A. rivasi (AMOVA: ΦCT = 0.420). The ARB + PUG barrier causes another break for A. rivasi (ΦCT = 0.406) and C. pica (ΦCT = 0.224). Three populations (K = 3) were identified for A. rivasi and C. pica, while N. tessellata presented one population (K = 1). The Mantel correlogram indicated that A. rivasi and C. pica fit the hierarchical population model, and only the A. rivasi and C. pica comparisons showed phylogeographic congruence. Our results demonstrate how the biological traits of these three species and the biogeographic barriers have influenced their phylogeographic structure.

Phylogenetic relationships of the Amblyomma cajennense complex (Acari: Ixodidae) at mitogenomic resolution.

The genus Amblyomma is the third most diverse in the number of species within the Ixodidae, with practically half of its species distributed in the Americas, though there are also species occurring in Africa, Asia, and Australia. Within the genus, there are several species complexes with veterinary and public health importance. The Amblyomma cajennense complex, in the Americas, is represented by six species with a wide distribution, from Texas to northern Argentina. We combined two sequencing techniques to generate complete mitogenomes of species belonging to the Amblyomma cajennense complex: genome skimming and long-range PCRs sequencing methods. Thus, we generated seven new mitochondrial genomes for all species of the Amblyomma cajennense complex, except for Amblyomma interandinum. Genetic distances between the mitogenomes corroborate the clear differentiation between the five species of the Amblyomma cajennense complex. The phylogenetic relationships of these species had previously been evaluated by combining partial nuclear and mitochondrial genes and here these relationships are corroborated with a more robust framework of data, which demonstrates that the conjunction of mitochondrial and nuclear partial genes can resolve close relationships when entire genes or genomes are unavailable. The gene order, structure, composition, and length are stable across these mitogenomes, and they share the general characteristics of Metastriata. Future studies should increase the number of available mitogenomes for this genus, especially for those species from the Indo-Pacific region and Africa, by means of a better understanding of their relationships and evolutionary process.

Molecular Detection of Rickettsia and Other Bacteria in Ticks and Birds in an Urban Fragment of Tropical Dry Forest in Magdalena, Colombia.

Birds are important hosts in the life cycle of some species of ticks. In Colombia, there are few eco-epidemiological studies of tick-borne diseases; the existing ones have been focused on areas where unusual outbreaks have occurred. This study describes the identification of ticks collected from birds and vegetation, and the detection of bacteria in those ticks and in blood samples from birds in an urban fragment of tropical dry forest in the department of Magdalena, Colombia. Bird sampling was carried out monthly in 2021, and 367 birds, distributed among 41 species, were captured. All collected ticks were identified as Amblyomma sp. or Amblyomma dissimile. The presence of rickettsiae in ticks collected from birds was evaluated by molecular analysis of the gltA, ompA and sca1 genes. 16S rRNA meta-taxonomy was used to evaluate rickettsiae in ticks collected from vegetation and in blood samples from birds. The presence of the species "Candidatus Rickettsia colombianensi" was detected in ticks from birds. Bacteria of the family Rickettsiacea was the most abundant in ticks collected from vegetation. Bacteria of the families Staphylococcaceae, Comamonadaceae and Pseudomonadaceae were prevalent in the samples of blood from birds. Rickettsia spp. was also detected in low abundance in some of the bird blood samples.

Molecular detection of Candidatus Rickettsia colombianensi in ticks (Acari, Ixodidae) collected from herpetofauna in San Juan de Carare, Colombia.

Knowledge about ticks and Rickettsiae in Colombia is still limited and the areas of the country where studies have been conducted are scarce. In this study, ticks (Acari, Ixodidae) associated with reptiles and amphibians in San Juan de Carare, Santander department, were morphologically and molecularly identified and tested for the presence of Rickettsia. For the molecular characterization of ticks, CO1, 12S and 16S sequences were generated and compared with other sequences available in genbank. Our analyses confirmed that the collected ticks were Amblyomma dissimile, and we provide the first report of this species parasitizing the snake Leptodeira septentrionalis. Of the samples analyzed, 69% were positive for Rickettsia sp. using the gltA, ompA and sca1 genes. Rickettsia sequences generated in this study were also compared to sequences downloaded from genbank by Bayesian inference (BI) and maximum likelihood (ML) phylogenetic analyzes. The presence of a single Rickettsia species, Candidatus Rickettsia colombianensi, was identified. This study expands the knowledge regarding the distribution of A. dissimile ticks and Rickettsiae in Colombia.

DNA barcoding reveals fraud in commercial common snook (Centropomus undecimalis) products in Santa Marta, Colombia.

The common snook Centropomus undecimalis is one of the main commercial fish species in the Caribbean region, including Colombia, where its populations have drastically decreased due to overfishing and environmental degradation. Thus, there is a market imbalance between the availability of snook products and their demand by consumers, which creates an opening for fraudulent actions such as species substitutions. Legislation in Colombia (and most Caribbean countries) lacks effective tools for the easy and rapid detection of frauds. Furthermore, there are very few studies published in scientific journals addressing this issue, of which none include C. undecimalis as the target species. Therefore, in order to investigate the existence of mislabeling in common snook products in Santa Marta, the present study analysed 44 frozen snook fillets from the five commercial brands available in the city. Moreover, 15 fresh snook fillets from six of the main fish markets were also analysed. To discover the frequency of possible frauds in labeling, samplings were carried out in July, September and November of 2019. Sample analyses involved the identification of each fillet at species level through molecular barcodes (16S-rRNA and COI), whose sequences were verified using BLAST and BOLD, and corroborated by a phylogenetic analysis. As a result, an astonishing 98% of the supermarkets fillets were found to be fraudulent, contrasting with a single case registered in the fish shop samples. The species used to substitute snook include the Pacific bearded brotula Brotula clarkae (38 samples), the Nile perch Lates niloticus (4 samples) and the acoupa weakfish Cynoscion acoupa (1 sample). Based on these results, there is a high rate of fraudulent labeling in the marketing of common snook in the city of Santa Marta, which calls for urgent actions to be taken by the corresponding authorities.

Molecular characterization of mosquitoes (Diptera: Culicidae) from the Colombian rainforest.

A few studies have carried out the taxonomic and molecular characterization of sylvatic mosquito species in Latin America, where some species have been incriminated as vectors for arboviruses and parasites transmission. The present study reports the molecular characterization of mosquito species in the Sierra Nevada de Santa Marta, a natural ecosystem in the Northern coast of Colombia. Manual capture methods were used to collect mosquitoes, and the specimens were identified via classical taxonomy. The COI marker was used for species confirmation, and phylogenetic analysis was performed using the neighbor-joining method, with the Kimura-2-Parameters model. Aedes serratus , Psorophora ferox , Johnbelkinia ulopus , Sabethes chloropterus , Sabethes cyaneus , Wyeomyia aporonoma , Wyeomyia pseudopecten , Wyeomyia ulocoma and Wyeomyia luteoventralis were identified. We assessed the genetic variability of mosquitoes in this area and phylogenetic reconstructions allowed the identification at the species level. Classical and molecular taxonomy demonstrated to be useful and complementary when morphological characteristics are not well preserved, or the taxonomic group is not represented in public molecular databases.

Molecular detection of pathogens in ticks associated with domestic animals from the Colombian Caribbean region.

Tick-borne diseases constitute a problem for livestock and public health. Given the socio-economic and environmental conditions of the Colombian Caribbean, ticks are particularly abundant, in turn exposing domestic animals and people in contact with them to such diseases. This study evaluates the presence of Babesia spp., Anaplasma spp., Coxiella spp. and Borrelia spp. in domestic animal ticks (Amblyomma mixtum, A. dissimile, Dermacentor nitens, Rhipicephalus sanguineus and R. microplus) by conventional PCR. Findings show a prevalence of 12.5% of Babesia, 0% of Borrelia, 39.4% of Anaplasma and 52.9% of Coxiella, whereas 6.2% of a total sample of 104 tick pools presented coinfections between Babesia and Anaplasma. Among the molecularly identified species are Ba. vogeli, Ba. bigemina and A. marginale, in addition to two Coxiella species-one being C. mudrowiae and the other similar to an undescribed endosymbiont of Rhipicephalus sp. It is necessary to evaluate the vector capacity of ticks such as A. mixtum, D. nitens and R. sanguineus in the transmission of A. marginale. Moreover, it is necessary to explore the role that bacteria of the genus Coxiella might have both in the health of humans and animals, and in the metabolism and reproduction of ticks. This is the first report on Babesia vogeli and B. bigemina in ticks from the Colombian Caribbean, representing a risk to animal and human health.

Characterization of the complete mitochondrial genome of Amblyomma ovale, comparative analyses and phylogenetic considerations.

In this study, we sequenced two complete mitochondrial genomes of Amblyomma ovale, a tick of public health importance. Sequencing two distinct individuals, the resulting mitochondrial genomes were 14,756 and 14,760 bp in length and maintained the same gene order previously reported in Amblyomma. These were combined with RNA-seq derived mitochondrial sequences from three additional species, Amblyomma aureolatum, Amblyomma maculatum, and Amblyomma moreliae, to carry out mitogenome comparative and evolutionary analyses against all previously published tick mitochondrial genomes. We described a derivative genome rearrangement that isolates Ixodes from the remaining Ixodidae and consists of both a reverse translocation as well as an event of Tandem Duplication Random Loss. Genetic distance analyses indicated that cox2, nd1, nd5, and 16S are good candidates for future population studies in A. ovale. The phylogenetic analyses corroborated the utility of complete mitochondrial genomes as phylogenetic markers within the group. This study further supplements the genome information available for Amblyomma and facilitates future evolutionary and population genetic studies within the genus.

Molecular detection and characterization of Anaplasma platys and Ehrlichia canis in dogs from northern Colombia.

Ehrlichia canis and Anaplasma platys are intracellular tick-transmitted bacteria that infect dogs; there is evidence for limited zoonotic potential as well. The prevalence of E. canis in Colombia has been evaluated in different regions; however little is known about the prevalence or distribution of A. platys. Neither pathogen has been studied in the Magdalena region, thus the purpose of our study was to assess the prevalence of these pathogens in dogs attending veterinary clinics from the cities of Santa Marta and Ciénaga, and to assess possible associated risk factors for infection. A. platys and E. canis infections in blood were evaluated by Taqman PCR assays. E. canis was detected in 26/170 (15.3%, 95% CI 10.4%-21.8%) and A. platys in 34/168 (20.2%, 95% CI 14.6%-27.3%) of all dogs tested. Eleven dogs (6.5%, 95% CI 3.4-11.7%) were coinfected with both pathogens. Sequencing results showed low diversity within E. canis and within A. platys strains, however a strain of E. canis detected in our study area is genetically distinct from strains reported in another city of Colombia. Our results suggest that for A. platys, Santa Marta dogs were at greater risk than Ciénaga dogs, and that purebred dogs were at slightly lower risk in both areas. The confirmation of these pathogens in northern Colombia should cause concern for the possible co-transmission of these agents to humans or animals in the region.

Molecular detection of Rickettsia spp., Anaplasma platys and Theileria equi in ticks collected from horses in Tayrona National Park, Colombia.

Horses are among the domestic animals that closely interact with humans and are highly parasitized by ticks, which are the primary vectors of zoonoses. As horses in Tayrona National Natural Park (PNNT) are used as a means of transporting goods, luggage and people, they are in constant contact with wild animals, workers and tourists from different countries. These factors increase the transmission risk of hemoparasites. The purpose of this study was to determine the presence of Rickettsia sp., Anaplasma sp., and Theileria sp., in horse ticks in this protected area using conventional PCR. We collected 343 ticks of genera Amblyomma, Rhipicephalus and Dermacentor. Of the 61 samples analyzed by PCR, 18 (29.5%) individuals were positive for Rickettsia sp., 15 (24.5%) for Anaplasma sp. and 4 (6.6%) for Theileria sp. This is the first report of these hemoparasite genera in ticks associated with horses in this preserved natural area, demonstrating the importance of additional studies on the presence and epidemiology of hemoparasites and their vectors in domestic and wild animals in conserved areas with a high flow of tourists.

Hemogregarine and Rickettsial infection in ticks of toads from northeastern Colombia.

The toads Rhinella spp. are in constant contact with humans and domestic animals and are commonly parasitized by ticks, which are also potential vectors of pathogenic microorganisms, such as apicomplexans and rickettsia. However, little is known about microorganisms associated with toad ticks. In this work, we molecularly evaluated the presence of Rickettsia spp. and hemogregarines in ticks of Rhinella horribilis and R. humboldti in the Colombian Caribbean, finding two different species of Rickettsia: the colombianensi strain and one close to R. bellii. In the case of hemogregarines, since only 18S gene sequences are available, it is difficult to define species and place them correctly in a phylogeny, but most of our samples show a 99% identity with Hemolivia stellata, while others identical to each other seem to form another clade within this genre. All collected ticks were identified as Amblyomma dissimile, representing the first time that H. stellata was recorded in this tick. The prevalence of both microorganisms was very high, which makes it necessary to generate robust phylogenies to clarify their taxonomic diversity and to correctly define their ecological role and pathogenicity, which should be taken into account in amphibian conservation plans and veterinary medicine.

Molecular identification of Giardia intestinalis in two cities of the Colombian Caribbean Coast.

INTRODUCTION: Parasitoses are an important health problem and its post-infectious consequences can cause high morbidity and mortality, worldwide. Epidemiological data is scarce in our region. We aimed to obtain frequency and circulating genotypes of Giardia intestinalis in two cities of the Caribbean Coast. METHODS: To identify parasites in samples, a fecal concentration and microscopical examination were performed. For further identification and genotyping of G. intestinalis, molecular analyses were performed using specific available primers. RESULTS: Only 22.1% of participants had gastrointestinal symptoms (most frequently: abdominal pain and diarrhea). Microscopy examination was positive in 72.7% of the samples. The most frequent parasite was G. intestinalis (48.1%) and we found mixed parasitic infections in 46.7% of the participants. All the samples from Santa Marta were molecularly characterized as assemblage A of G. intestinalis, while in Cartagena we confirmed the presence of assemblages A and B. CONCLUSIONS: Molecular tools are important in the diagnosis of parasites and to increase comprehension of their distribution. In this study, a higher frequency of parasites, mostly protozoa, was observed.

Rickettsial infection in ticks (Acari: Ixodidae) from reptiles in the Colombian Caribbean.

Although more reptiles are illegally traded in Colombia than any other group of animals, for both local and international markets, little is known about ticks associated with reptiles or pathogens associated with these ticks. In this study, ticks were collected from reptiles in Magdalena, Cesar and La Guajira regions in northern Colombia, and identified morphologically using taxonomic keys and molecularly by sequencing of the COI gene. In addition, Rickettsia spp. were detected by PCR amplifying the gltA, 16S rRNA, and sca1 genes. Ticks were identified as Amblyomma dissimile, representing the first record for this species in 17 of the 26 species of reptiles sampled. Additionally, our data expand the distribution range for this tick in Colombia. Rickettsia spp. DNA was detected in 18 ticks (3.6%) from 16 reptiles. Sequence analyses indicated that most of the Rickettsia were members of the R. monacensis clade, most closely related to Rickettsia sp. strain Colombianensi. We detected one tick infected with a Rickettsia with 99% identity to R. bellii.

Nipponnemertes incainca sp. n. Adoption of the new taxonomic proposal for nemerteans (Nemertea, Cratenemertidae).

A new species Nipponemertes incainca is described from the intertidal zone of Santa Marta, Colombia. A new recent approach based on both morphological and molecular characters is applied for the description. The main characteristics of the species are: red color, head shield-shaped with a mid-dorsal cephalic ridge, furrows pre-cerebral inconspicuous with few faint ridges orthogonal to furrow axis, two irregular groups of eyespots situated at lateral margins in precerebral cephalic region, proboscis provided with papillae and 12 nerves, stylet smooth supported on an oval basis, and two pouches containing 3-4 accessory stylets each. The sequence of the COI gene was analyzed as an additional support for the new species.

Its2 para la identificación de califóridos (diptera: calliphoridae) de importancia forense en colombia / ITS2 for the Identification of Calliphoridae (Diptera: Calliphoridae) of Forensic Importance in Colombia

La entomología forense es una disciplina que utiliza insectos para obtener información útil en la determinación del intervalo postmortem (IPM). Las moscas de la familia Calliphoridae son muy utilizadas en entomología forense, sin embargo, su identificación a nivel de especie puede dificultarse cuando el individuo se encuentra incompleto o en estadio inmaduro. En el presente trabajo, se evaluó el potencial de la región ITS2 del genoma nuclear para la identificación de especies de Calliphoridae en Colombia utilizando tres aproximaciones: comparando distancias genéticas utilizando la metodología de códigos de barra, haciendo una reconstrucción filogenética, y con enzimas de restricción (PCR-RFLPs). Se secuenciaron un total de 520 pb en 44 individuos pertenecientes a 16 especies. Se calcularon los valores de distancia intraespecífica e interespecíficas utilizando el modelo K2P. Los valores de distancia intraespecífica oscilaron entre 0 y 0,252 %, mientras que las distancias interespecíficas fluctuaron entre 3,6 y 18,9 %, evidenciándose que esta técnica puede ser utilizada como código de barras genético en la identificación de especies de la familia Calliphoridae. Tanto en los análisis de Neighbour-Joining como en los análisis bayesianos el 90 % de los géneros presentan una monofilia sustentada en probabilidad posterior de 0,89 a 1. En todos los casos la especie Blepharicnema splendens agrupa con el género Lucilia. Con base en las secuencias obtenidas se utilizó la aplicación NEBCutter para identificar cuatro enzimas de restricción las cuales se probaron en el laboratorio y se comprobó su utilidad para la identificación rápida de especies de Calliphoridae en Colombia.

Forensic entomology is a discipline that uses insects to obtain useful information for the determination of the postmortem interval (PMI). Flies of the family Calliphoridae are extensively used for this purpose, however, the identification of these flies can be difficult when the individual is not an adult or when it is incomplete. In the present work, we tested the utility of the ITS2 region of the nuclear genome for the identification of Calliphoridae species in Colombia using three approaches: comparing genetic distances using the barcoding methodology, with a phylogenetic reconstruction, and with PCR-RFLPs. We sequenced 520 bp in 44 individuals belonging to 16 species of califorids. Intraspecific and interspecific distance values were calculated using the K2P model. The intraspecific distance values ranged between 0 and 0.252 %, while the interspecific distance values ranged between 3.6 and 18.9 %, indicating that this gene can be used as a genetic barcode for the identification of species of the Calliphoridae family. Both the Neighbour-Joining and Bayesian analyses recovered 90 % of the genera as monophyletic, with pp values between 0.89 and 1. Blepharicnema splendens was always recovered within the Lucilia genera. Based on the obtained sequences we used the NEBCutter application to identify four restriction enzymes that cut in a differential way and generated useful patterns for the identification of the species. The enzymes were successfully tested and confirmed the utility of this technique as a fast way to identify species of Calliphoridae in Colombia.

Phylogenetic relationships among superfamilies of Neritimorpha (Mollusca: Gastropoda).

Despite the extraordinary morphological and ecological diversity of Neritimorpha, few studies have focused on the phylogenetic relationships of this lineage of gastropods, which includes four extant superfamilies: Neritopsoidea, Hydrocenoidea, Helicinoidea, and Neritoidea. Here, the nucleotide sequences of the complete mitochondrial genomes of Georissa bangueyensis (Hydrocenoidea), Neritina usnea (Neritoidea), and Pleuropoma jana (Helicinoidea) and the nearly complete mt genomes of Titiscania sp. (Neritopsoidea) and Theodoxus fluviatilis (Neritoidea) were determined. Phylogenetic reconstructions using probabilistic methods were based on mitochondrial (13 protein coding genes and two ribosomal rRNA genes), nuclear (partial 28S rRNA, 18S rRNA, actin, and histone H3 genes) and combined sequence data sets. All phylogenetic analyses except one converged on a single, highly supported tree in which Neritopsoidea was recovered as the sister group of a clade including Helicinoidea as the sister group of Hydrocenoidea and Neritoidea. This topology agrees with the fossil record and supports at least three independent invasions of land by neritimorph snails. The mitochondrial genomes of Titiscania sp., G. bangueyensis, N. usnea, and T. fluviatilis share the same gene organization previously described for Nerita mt genomes whereas that of P. jana has undergone major rearrangements. We sequenced about half of the mitochondrial genome of another species of Helicinoidea, Viana regina, and confirmed that this species shares the highly derived gene order of P. jana.

Molecular detection of Rickettsia, Borrelia, and Babesia species in Ixodes ricinus sampled in northeastern, central, and insular areas of Italy.

The aim of the present study was to provide insight into the diversity of tick-borne pathogens circulating in Italy, carried/transmitted by Ixodes ricinus, one of the most abundant tick species in the country. A total of 447 specimens sampled in five areas of northeastern, central and insular Italy were analysed by polymerase chain reaction and sequencing for the presence of rickettsiae, borreliae and babesiae. Several rickettsial species of the spotted fever group of zoonotic concern and other zoonotic pathogens were found, such as Borrelia burgdorferi s.s., Borrelia afzelii, Borrelia garinii, and Babesia venatorum. These findings confirm a wide distribution of tick-borne bacterial and protozoan species in Italy, and highlight the sanitary importance of I. ricinus, often recorded as feeding on humans.